PT1342410E - Use of a galanin agonist in the preparation of a medicament for improving memory and other cognitive functions - Google Patents
Use of a galanin agonist in the preparation of a medicament for improving memory and other cognitive functions Download PDFInfo
- Publication number
- PT1342410E PT1342410E PT02028584T PT02028584T PT1342410E PT 1342410 E PT1342410 E PT 1342410E PT 02028584 T PT02028584 T PT 02028584T PT 02028584 T PT02028584 T PT 02028584T PT 1342410 E PT1342410 E PT 1342410E
- Authority
- PT
- Portugal
- Prior art keywords
- galanin
- gene
- treatment
- mice
- mouse
- Prior art date
Links
- 101800002068 Galanin Proteins 0.000 title claims abstract description 59
- SLZIZIJTGAYEKK-CIJSCKBQSA-N molport-023-220-247 Chemical compound C([C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)CN)[C@@H](C)O)C1=CNC=N1 SLZIZIJTGAYEKK-CIJSCKBQSA-N 0.000 title claims abstract description 56
- 102000019432 Galanin Human genes 0.000 title claims abstract description 54
- 239000000556 agonist Substances 0.000 title claims abstract description 9
- 230000003920 cognitive function Effects 0.000 title claims description 5
- 239000003814 drug Substances 0.000 title claims description 4
- 238000002360 preparation method Methods 0.000 title claims description 4
- 208000024827 Alzheimer disease Diseases 0.000 claims abstract description 8
- 210000005036 nerve Anatomy 0.000 abstract description 10
- 230000008929 regeneration Effects 0.000 abstract description 10
- 238000011069 regeneration method Methods 0.000 abstract description 10
- 239000005557 antagonist Substances 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 8
- 208000002193 Pain Diseases 0.000 abstract description 3
- 230000036407 pain Effects 0.000 abstract description 3
- 208000005877 painful neuropathy Diseases 0.000 abstract description 3
- 206010002091 Anaesthesia Diseases 0.000 abstract 1
- 208000028389 Nerve injury Diseases 0.000 abstract 1
- 206010036832 Prolactinoma Diseases 0.000 abstract 1
- 230000037005 anaesthesia Effects 0.000 abstract 1
- 238000001949 anaesthesia Methods 0.000 abstract 1
- 230000019771 cognition Effects 0.000 abstract 1
- 238000011813 knockout mouse model Methods 0.000 abstract 1
- 230000008764 nerve damage Effects 0.000 abstract 1
- 208000030153 prolactin-producing pituitary gland adenoma Diseases 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 22
- 108090000623 proteins and genes Proteins 0.000 description 12
- 241001465754 Metazoa Species 0.000 description 10
- 210000002569 neuron Anatomy 0.000 description 10
- 241000699666 Mus <mouse, genus> Species 0.000 description 9
- 230000027928 long-term synaptic potentiation Effects 0.000 description 8
- 210000004556 brain Anatomy 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 210000001320 hippocampus Anatomy 0.000 description 6
- 230000035772 mutation Effects 0.000 description 6
- 230000008685 targeting Effects 0.000 description 6
- 108700024394 Exon Proteins 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 230000007423 decrease Effects 0.000 description 5
- 230000001817 pituitary effect Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 4
- 210000003050 axon Anatomy 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 210000001671 embryonic stem cell Anatomy 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 230000002779 inactivation Effects 0.000 description 4
- 208000014674 injury Diseases 0.000 description 4
- 230000013011 mating Effects 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 210000003497 sciatic nerve Anatomy 0.000 description 4
- 230000001953 sensory effect Effects 0.000 description 4
- 241001282736 Oriens Species 0.000 description 3
- 241000283984 Rodentia Species 0.000 description 3
- 238000002105 Southern blotting Methods 0.000 description 3
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 210000003016 hypothalamus Anatomy 0.000 description 3
- 230000011514 reflex Effects 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 208000000044 Amnesia Diseases 0.000 description 2
- 108010058699 Choline O-acetyltransferase Proteins 0.000 description 2
- 102100023460 Choline O-acetyltransferase Human genes 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 208000026139 Memory disease Diseases 0.000 description 2
- 101500025322 Rattus norvegicus Galanin Proteins 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 229960004373 acetylcholine Drugs 0.000 description 2
- 230000036592 analgesia Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 206010012601 diabetes mellitus Diseases 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 230000000971 hippocampal effect Effects 0.000 description 2
- -1 i. e.g. Proteins 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 230000030214 innervation Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000006984 memory degeneration Effects 0.000 description 2
- 208000023060 memory loss Diseases 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 210000001428 peripheral nervous system Anatomy 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 210000001044 sensory neuron Anatomy 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 210000003594 spinal ganglia Anatomy 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 230000008733 trauma Effects 0.000 description 2
- SFLSHLFXELFNJZ-QMMMGPOBSA-N (-)-norepinephrine Chemical compound NC[C@H](O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-QMMMGPOBSA-N 0.000 description 1
- 229930000680 A04AD01 - Scopolamine Natural products 0.000 description 1
- 108700028369 Alleles Proteins 0.000 description 1
- 101000583080 Bunodosoma granuliferum Delta-actitoxin-Bgr2a Proteins 0.000 description 1
- 208000000094 Chronic Pain Diseases 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 208000025962 Crush injury Diseases 0.000 description 1
- 206010012289 Dementia Diseases 0.000 description 1
- 208000017701 Endocrine disease Diseases 0.000 description 1
- 101710169265 Galanin peptides Proteins 0.000 description 1
- 102100028501 Galanin peptides Human genes 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000018997 Growth Hormone Human genes 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- 208000009889 Herpes Simplex Diseases 0.000 description 1
- 101000860415 Homo sapiens Galanin peptides Proteins 0.000 description 1
- STECJAGHUSJQJN-GAUPFVANSA-N Hyoscine Natural products C1([C@H](CO)C(=O)OC2C[C@@H]3N([C@H](C2)[C@@H]2[C@H]3O2)C)=CC=CC=C1 STECJAGHUSJQJN-GAUPFVANSA-N 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 101500025319 Mus musculus Galanin Proteins 0.000 description 1
- STECJAGHUSJQJN-UHFFFAOYSA-N N-Methyl-scopolamin Natural products C1C(C2C3O2)N(C)C3CC1OC(=O)C(CO)C1=CC=CC=C1 STECJAGHUSJQJN-UHFFFAOYSA-N 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 102000003797 Neuropeptides Human genes 0.000 description 1
- 108090000189 Neuropeptides Proteins 0.000 description 1
- 206010034620 Peripheral sensory neuropathy Diseases 0.000 description 1
- 206010034719 Personality change Diseases 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 102000005157 Somatostatin Human genes 0.000 description 1
- 108010056088 Somatostatin Proteins 0.000 description 1
- 108700009124 Transcription Initiation Site Proteins 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 229940035676 analgesics Drugs 0.000 description 1
- 239000000730 antalgic agent Substances 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000003542 behavioural effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000002459 blastocyst Anatomy 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 210000003710 cerebral cortex Anatomy 0.000 description 1
- 108091006116 chimeric peptides Proteins 0.000 description 1
- 230000001713 cholinergic effect Effects 0.000 description 1
- 210000002932 cholinergic neuron Anatomy 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000003930 cognitive ability Effects 0.000 description 1
- 230000001149 cognitive effect Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 229960003638 dopamine Drugs 0.000 description 1
- 210000001198 duodenum Anatomy 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 229960003692 gamma aminobutyric acid Drugs 0.000 description 1
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 102000050963 human GAL Human genes 0.000 description 1
- 208000008384 ileus Diseases 0.000 description 1
- 238000003365 immunocytochemistry Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 230000003914 insulin secretion Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 208000028774 intestinal disease Diseases 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000007087 memory ability Effects 0.000 description 1
- 230000006386 memory function Effects 0.000 description 1
- CWWARWOPSKGELM-SARDKLJWSA-N methyl (2s)-2-[[(2s)-2-[[2-[[(2s)-2-[[(2s)-2-[[(2s)-5-amino-2-[[(2s)-5-amino-2-[[(2s)-1-[(2s)-6-amino-2-[[(2s)-1-[(2s)-2-amino-5-(diaminomethylideneamino)pentanoyl]pyrrolidine-2-carbonyl]amino]hexanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-5 Chemical compound C([C@@H](C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)OC)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CCCN=C(N)N)C1=CC=CC=C1 CWWARWOPSKGELM-SARDKLJWSA-N 0.000 description 1
- 229960005181 morphine Drugs 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000004898 n-terminal fragment Anatomy 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 230000007472 neurodevelopment Effects 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 229960002748 norepinephrine Drugs 0.000 description 1
- SFLSHLFXELFNJZ-UHFFFAOYSA-N norepinephrine Natural products NCC(O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-UHFFFAOYSA-N 0.000 description 1
- 231100000862 numbness Toxicity 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 210000002963 paraventricular hypothalamic nucleus Anatomy 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- STECJAGHUSJQJN-FWXGHANASA-N scopolamine Chemical compound C1([C@@H](CO)C(=O)O[C@H]2C[C@@H]3N([C@H](C2)[C@@H]2[C@H]3O2)C)=CC=CC=C1 STECJAGHUSJQJN-FWXGHANASA-N 0.000 description 1
- 229960002646 scopolamine Drugs 0.000 description 1
- 201000005572 sensory peripheral neuropathy Diseases 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- NHXLMOGPVYXJNR-ATOGVRKGSA-N somatostatin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CSSC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N1)[C@@H](C)O)NC(=O)CNC(=O)[C@H](C)N)C(O)=O)=O)[C@H](O)C)C1=CC=CC=C1 NHXLMOGPVYXJNR-ATOGVRKGSA-N 0.000 description 1
- 229960000553 somatostatin Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000000278 spinal cord Anatomy 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
- A01K67/0276—Knock-out vertebrates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/07—Animals genetically altered by homologous recombination
- A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/0306—Animal model for genetic diseases
- A01K2267/0312—Animal model for Alzheimer's disease
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/0306—Animal model for genetic diseases
- A01K2267/0318—Animal model for neurodegenerative disease, e.g. non- Alzheimer's
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/035—Animal model for multifactorial diseases
- A01K2267/0356—Animal model for processes and diseases of the central nervous system, e.g. stress, learning, schizophrenia, pain, epilepsy
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Organic Chemistry (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Diabetes (AREA)
- Endocrinology (AREA)
- General Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Environmental Sciences (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Animal Husbandry (AREA)
- Obesity (AREA)
- Molecular Biology (AREA)
- Plant Pathology (AREA)
- Microbiology (AREA)
- Hematology (AREA)
Abstract
Description
DESCRIÇÃO "UTILIZAÇÃO DE UM AGONISTA DE GALANINA NA PREPARAÇÃO DE UM MEDICAMENTO PARA MELHORAR A MEMÓRIA E OUTRAS FUNÇÕES COGNITIVAS" A invenção refere-se à galanina, incluindo os seus análogos e suas utilizações. A galanina é um neuropéptido de 29 aminoácidos que foi isolado pela primeira vez a partir do intestino do porco, em 1983. Depois disso fez-se a clonagem do ADNc da galanina a partir de um banco da pituitária anterior do rato, em 1987. A análise das sequências de nucleótidos e de aminoácidos sugere que a galanina não está relacionada com nenhuma das outras familias conhecidas de péptidos reguladores, continuando a ser o único membro desta familia. A porção do terminal N da galanina é altamente conservada entre espécies, havendo uma variação na porção do terminal C. A galanina está profusamente distribuída nos sistemas nervosos periférico e central, no intestino e no pâncreas. Existe em concentrações elevadas na saliência mediana do hipotálamo e na pituitária. 0 documento W092/12997 (General Hospital Corporation), publicado em 1992, divulga a sequência da galanina humana. Estão a ser analisados estudos de outros autores que prevêem a administração da galanina do rato ou dos seus fragmentos do terminal N, para aumentar o efeito da morfina, sugerindo esse pedido de patente de invenção que é previsível que a galanina 1 tenha efeitos analgésicos tais que possa ser administrada por si só ou em combinação com outros analgésicos. 0 pedido reivindica a utilização da galanina ou dos seus análogos, para o tratamento da dor e a utilização de antagonistas da galanina para o tratamento de outras patologias. 0 documento W092/20709 (Astra AB) divulga diversos antagonistas putativos da galanina. Os antagonistas que são ali descritos têm todos por base os primeiros 12 aminoácidos da galanina, a que se seguem sequências parciais de outros péptidos, i. e., péptidos quiméricos. Alguns podem ser agonistas, outros podem ser antagonistas e outros ainda podem ser de ambas, dependendo do subtipo de receptor. 0 pedido divulga que os antagonistas podem ser úteis para o tratamento de patologias associadas à insulina, à hormona do crescimento, à acetil-colina, à dopamina, à Substância P, à Somatostatina e à noradrenalina, incluindo as doenças do foro endocrinológico, ingestão alimentar, neurológico e psiquiátrico, demência de tipo Alzheimer, analgesias, doenças intestinais. 0 pedido divulga os resultados de estudos em que são utilizados alguns dos antagonistas ali descritos para se obter diversos efeitos, tais como a inibição, determinada pela galanina, da libertação de insulina estimulada pela glucose; a inibição, induzida pela galanina, da libertação da Ach do hipocampo induzida pela escopolamina; a facilitação, induzida pela galanina, do reflexo flexor; e o deslocamento da galanina iodada ligada, em estudos de ligação membranares. Há uma sugestão nesse pedido, em que se admite que os antagonistas possam ser prescritos para analgesia, mas no pedido não há nenhuma divulgação relativa a este efeito.Use of a galanine agonist in the preparation of a medicament for improving memory and other cognitive functions. The invention relates to galanin, including analogues thereof and uses thereof. Galanin is a 29 amino acid neuropeptide which was first isolated from the gut of the pig in 1983. Thereafter the cDNA from galanin was cloned from a rat anterior pituitary bank in 1987. The galanin cDNA analysis of nucleotide and amino acid sequences suggests that galanin is unrelated to any of the other known families of regulatory peptides, remaining the only member of this family. The N-terminal portion of galanin is highly conserved between species, with a variation on the C-terminal portion. Galanin is widely distributed in the peripheral and central nervous systems, the intestine and the pancreas. It exists in high concentrations in the medial protrusion of the hypothalamus and pituitary. W092 / 12997 (General Hospital Corporation), published in 1992, discloses the human galanin sequence. Studies of other authors are contemplated which provide for the administration of rat galanin or its N-terminal fragments to increase the effect of morphine, suggesting such a patent application that it is expected that galanin 1 will have analgesic effects such that be administered alone or in combination with other analgesics. The application claims the use of galanin or its analogues for the treatment of pain and the use of galanin antagonists for the treatment of other conditions. W092 / 20709 (Astra AB) discloses several putative galanin antagonists. The antagonists which are described therein are all based on the first 12 amino acids of galanin, which are followed by partial sequences of other peptides, i. e.g., chimeric peptides. Some may be agonists, others may be antagonists and still others may be of both, depending on the receptor subtype. The application discloses that the antagonists may be useful for the treatment of conditions associated with insulin, growth hormone, acetylcholine, dopamine, Substance P, Somatostatin and noradrenaline, including endocrine disorders, food intake , neurological and psychiatric, Alzheimer's type dementia, analgesia, intestinal diseases. The application discloses the results of studies in which some of the antagonists described therein are used to achieve various effects, such as the inhibition, determined by galanin, of glucose-stimulated insulin release; the galanin-induced inhibition of scopolamine-induced hippocampus Ach release; the galanin-induced facilitation of the flexor reflex; and the displacement of bound iodinated galanin in membrane binding studies. There is a suggestion in this application, where it is accepted that antagonists may be prescribed for analgesia, but in the application there is no disclosure relating to this effect.
Aproximadamente 2-4% da população Ocidental padece de diabetes mellitus e entre estas pessoas há 10-15% que padecem de 2 dores crónicas e dormência nas suas extremidades, designada por "neuropatia dolorosa". As técnicas actuais para o combate à neuropatia dolorosa são inadequadas. A doença de Alzheimer é uma causa muito importante de morbidade em todo o mundo, sendo esta doença caracterizada pela perda de memória e por alterações da personalidade. Ao nível anatómico há uma diminuição muito significativa do número de nervos colinérgicos no córtex cerebral basal e no hipocampo, que são as zonas principais do cérebro que, presumivelmente, armazenam dados em memória e com eles executam operações. Trabalhos anteriores comprovaram que a galanina também é expressa nestes nervos do hipocampo e que as concentrações de galanina são duas vezes mais elevadas nos cérebros de doentes com doença de Alzheimer. A invenção refere-se a um método para melhorar a memória, aumentar a memória e melhorar a função cognitiva, compreendendo a administração de um agonista de galanina a um indivíduo. De um modo vantajoso, tal tratamento pode ser utilizado no tratamento da restauração da memória após lesão, trauma ou na doença de Alzheimer. A presente invenção proporciona a utilização de um agoniste de galanina na preparação de um medicamento para o tratamento da doença de Alzheimer e doenças e estados relacionados, e para aumentar a memória e a função cognitiva.Approximately 2-4% of the Western population suffers from diabetes mellitus and among these people there are 10-15% who suffer from 2 chronic pains and numbness in their extremities, called " painful neuropathy ". Current techniques for combating painful neuropathy are inadequate. Alzheimer's disease is a very important cause of morbidity around the world, being this disease characterized by memory loss and personality changes. At the anatomical level there is a very significant decrease in the number of cholinergic nerves in the basal cerebral cortex and the hippocampus, which are the major zones of the brain that presumably store data in memory and perform operations with it. Previous work has shown that galanin is also expressed in these hippocampal nerves and that galanin concentrations are twice as high in the brains of patients with Alzheimer's disease. The invention relates to a method for improving memory, enhancing memory and improving cognitive function, comprising administering a galanin agonist to a subject. Advantageously, such treatment can be used in the treatment of memory restoration after injury, trauma or in Alzheimer's disease. The present invention provides the use of a galanin agonist in the preparation of a medicament for the treatment of Alzheimer's disease and related diseases and conditions, and for enhancing memory and cognitive function.
Seguidamente serão descritas formas de realização da invenção apenas a título exemplificativo, com referência os desenhos anexos, as Figuras 1 a 9 nas quais: 3 a Fig. 1 ilustra a estrutura genómica da galanina do murganho, a Fig.2 ilustra o vector de direccionamento utilizado para a produção do roedor da presente invenção, a Fig.3 ilustra o evento de recombinação específico na produção do roedor em conformidade com a invenção, a Fig.4 ilustra o genotipo da progenia, determinado utilizando a metodologia de análise por transferênciade Southern e por PCR, demonstrando a existência de resultados idênticos a partir da mesma ninhada proveniente de um acasalamento de dois animais heterozigóticos, a Fig.a 5 ilustra efeito da inactivação da galanina sobre a regeneração de curto prazo dos neurónios sensoriais, a Fig.6 ilustra o efeito da inactivação da galanina sobre a regeneração de longo prazo de neurónios sensoriais, a Fig.7 ilustra a expressão de uma ribossonda específica do exão 6 para se estudar a distribuição dos neurónios galaninérgicos no cérebro e no gânglio da raiz dorsal de murganhos de tipo natural e de murganhos mutantes, a Fig.8 ilustra os efeitos da inactivação da galanina sobre a geração de uma potenciação de longo prazo na zona do stratum radiatum do hipocampo; e a Fig.9 ilustra os efeitos da inactivação da galanina sobre a geração da potenciação de longo prazo na zona do stratum oriens do hipocampo. 4Embodiments of the invention will now be described by way of example only, with reference to the accompanying drawings, Figures 1 to 9 in which: Fig. 1 illustrates the genomic structure of mouse galanin, Fig. 2 illustrates the targeting vector used for the production of the rodent of the present invention, Fig. 3 illustrates the specific recombination event in rodent production in accordance with the invention, Fig.4 illustrates progeny genotype, determined using Southern blot analysis methodology and by PCR, demonstrating the existence of identical results from the same litter from a mating of two heterozygous animals, Fig. 5 illustrates the inactivation effect of galanin on the short-term regeneration of sensory neurons, Fig. effect of galanin inactivation on long-term regeneration of sensory neurons, Fig. 7 illustrates the expression of a ribos Fig. 8 illustrates the effects of inactivation of galanin on the generation of long-term potentiation of galaninergic neurons in the brain and dorsal root ganglion of wild-type mice and mutant mice. term in the hippocampus stratum radiatum zone; and Fig. 9 illustrates the effects of inactivation of galanin on the generation of long-term potentiation in the hippocampus stratum oriens zone. 4
Para se gerar uma mutação num murganho, isto é, para se introduzir num genoma de um murganho quer uma mutação de um locus de um gene especifico com perda de função ou com ganho de função (de acordo com o processo descrito por Kuehn, M. R. et al., Nature, 1987; 326:295-8; Thomas, K. R. e Capecchi, M. R. em Nature, 1986; 324:34-8), são necessários vários passos: (1) a clonagem do locus relevante genómico do murganho; (2) a construção de um vector de direccionamento, de tal modo que o locus/gene de interesse seja modificado para inactivar ou alterar a sua estrutura e a sua função de alguma forma; (3) a introdução do vector de direccionamento num banco de células estaminais embrionárias e a selecção e a identificação de clones de células singulares onde o evento de direccionamento adequado, correcto tenha tido lugar e onde tenha ficado inalterado o número de cromossomas; e (4} a introdução desses clones em blastócitos com 3,5 dias de idade com o acasalamento dos murganhos quiméricos resultantes com tipos naturais do sexo oposto. A descendência resultante demonstrou ser portadora da mutação, pelo que são assim seres heterozigóticos e graças a um acasalamento adequado são criados seres homozigóticos relativamente à mutação introduzida.To generate a mutation in a mouse, i.e. to introduce into a mouse genome either a mutation of a specific gene with loss of function or gain of function (according to the procedure described by Kuehn, M. et al. (1) the cloning of the relevant genomic locus of the mouse is required; (1) cloning of the relevant genomic locus of the mouse; (2) constructing a targeting vector such that the locus / gene of interest is modified to inactivate or alter its structure and function in some way; (3) introduction of the targeting vector into a bank of embryonic stem cells and selection and identification of single cell clones where the appropriate, correct targeting event has taken place and where the number of chromosomes has remained unchanged; and (4) the introduction of such clones into 3.5 days old blastocysts by mating the resulting chimeric mice with natural types of the opposite sex. The resulting offspring were shown to carry the mutation, thus being heterozygous and mating are created homozygous beings with respect to the introduced mutation.
Como um primeiro passo efectuou-se a clonagem do gene da galanina dos murinos. Efectuou-se uma pesquisa da biblioteca genómica de murganhos (Ehrich, E. et al., Gene, 1987; 57:229-37) utilizando como uma sonda o ADNc de galanina de rato com o comprimento completo, em condições de elevado rigor. Foram identificados dois clones cosmidicos com uma extensão de 60 kB em torno do locus da galanina. Utilizando sondas de 5' e de 3', obtidas a partir do ADNc do rato, efectuou-se a subclonagem de uma região de 14 Kb de ADN que continha o gene completo, tendo 5 sido feita a sua sequenciação parcial. A partir da sequência genómica foram concebidos iniciadores complementares para as regiões exónicas não traduzidas do gene. Foi gerado um fragmento de 630 pb por PCR-TR (kit fornecido por INVITROGEN BV, Holanda), utilizando como fonte de ARNm o cérebro intacto de uma fêmea adulta. A sequenciação subsequente deste fragmento demonstrou que as galaninas do murganho e do rato são 100% idênticas ao nível das proteínas e 94,8% idênticas ao nível dos nucleótidos. A estrutura genómica do gene do murganho (Fig. 1} é idêntica à do gene do rato. O gene abrange 4,8 Kb e consiste em seis exões. O local de início da tradução (AUG) começa na primeira base do segundo exão e a região codificadora para a galanina desenvolve-se através dos exões, terceiro e quarto ficando o codão de paragem (UGA) no meio do sexto exão.As a first step the murine galanin gene was cloned. The genomic library of mice (Ehrich, E. et al., Gene, 1987; 57: 229-37) was screened using the full-length rat galanin cDNA under high stringency conditions as a probe. Two cosmid clones with a 60 kB extension were identified around the galanin locus. Using 5 'and 3' probes obtained from the mouse cDNA, a 14 kb region of DNA containing the complete gene was subcloned and partial sequencing done. From the genomic sequence complementary primers were designed for the non-translated exonic regions of the gene. A 630 bp fragment was generated by TR-PCR (kit provided by INVITROGEN BV, The Netherlands), using as the source of mRNA the intact brain of an adult female. Subsequent sequencing of this fragment demonstrated that mouse and rat gallanins are 100% protein-identical and 94.8% nucleotide-level. The genomic structure of the mouse gene (Fig. 1) is identical to that of the mouse gene. The gene covers 4.8 kb and consists of six exons. The translation start site (AUG) begins at the first base of the second exon and the coding region for galanin develops through the exons, third and fourth, leaving the stop codon (UGA) in the middle of the sixth exon.
Utilizando o subclone de 14 Kb descrito acima, construiu-se um vector de direccionamento para selecção positiva/negativa (Fig. 2) . A mutação introduzida remove os primeiros cinco exões que contêm a região codificadora completa do péptido de galanina (Fig. 3) .Using the 14 Kb subclone described above, a targeting vector for positive / negative selection was constructed (Fig. 2). The introduced mutation removes the first five exons that contain the complete coding region of the galanin peptide (Fig. 3).
Na figura 3, A e B são os locais das sondas externas utilizadas para a pesquisa das células de ES para a integração adequada do arquétipo.In Figure 3, A and B are the sites of the external probes used for the screening of ES cells for proper integration of the archetype.
Neo = gene de resistência à neomicinaNeo = neomycin resistance gene
VHS-TK = vírus do herpes simplex-gene da timidina-cinase B = Bamhl E = EcoRI X = Xhol Bg = BglII 6VHS-TK = herpes simplex virus-thymidine kinase gene B = Bamhl E = EcoRI X = XhoI Bg = BglII 6
Em particular, o gene de direccionamento remove um segmento de 3,2 Kb do ADN e, conseguentemente, remove os primeiros 5 exões do gene da galanina. Os locais exactos que flanqueiam o segmento de ADN removido são o local de S' do BamRI que fica 10 pb a jusante do local de inicio da transcrição e o local de 3' é o local de Bgll no meio do intrão 5. Estes locais estão indicados com asteriscos na Fig. 3. Outros locais que poderiam ser utilizados são o mesmo local de 5' e um local diferente de Xhol de 3' no intrão 4 que poderia remover apenas 2,9 Kb do ADN e poderia, por isso, remover apenas os primeiros 4 exões.In particular, the targeting gene removes a 3.2 kb segment of DNA and, accordingly, removes the first 5 exons of the galanin gene. The exact locations flanking the removed DNA segment are the BamRI S 'site which is 10 bp downstream from the transcription start site and the 3' site is the BgII site in the middle of the intron 5. These sites are indicated with asterisks in Fig. 3. Other sites that could be used are the same 5 'site and a different 3' Xho I site in intron 4 that could remove only 2.9 Kb from the DNA and could therefore remove only the first 4 exons.
Este vector foi linearizado e submetido a uma operação de electroporação na linhagem de células estaminais embrionárias (EE) E14 (Hooper, M. et al., Nature, 1987, 326:292-5). A cartografia de restrição do locus de tipo natural com BglII gera um fragmento de 9,3 Kb quando hibridado com uma sonda externa de 5' (marcada com A na Fig. 3), ao passo que o locus correctamente identificado gera um fragmento de 4,4 Kb. No total, foramThis vector was linearized and subjected to an electroporation step in the E14 embryonic stem cell (EE) lineage (Hooper, M. et al., Nature, 1987, 326: 292-5). Restriction mapping of the wild-type BglII locus generates a 9.3 Kb fragment when hybridized with a 5 'external probe (labeled with A in Fig. 3), whereas the correctly identified locus generates a fragment of 4 , 4 Kb. In total,
identificados 9 clones nos quais um alelo do gene da galanina foi correctamente identificado por recombinação homóloga entre 209 colónias duplamente resistentes, determinando uma frequência de marcação de 4,3%. Estes nove clones foram cariotipifiçados, confirmaram a euploidia e foram injectados em blastócitos com 3,5 dias de idade de murganhos da estirpe C57/BL/6. A transmissão da linhagem germinativa do locus da galanina destruído foi obtida a partir de três clones separados de células EE. Determinou-se o genotipo da progenia utilizando a metodologia de análise por transferência de Southern e por PCR (a Fig. 4 demonstra resultados idênticos obtidos pela metodologia de análise por transferência de Southern e por pesquisa por PCR na mesma ninhada obtida a partir de um acasalamento de dois animais heterozigóticos). A mutação 7 prosseguiu com uma procriação até â homozigoticidade na estirpe de serotipo-129 sv e todos os dados apresentados são do murganho com esta formação. 1. Os resultados da análise dos genotipos dos nascituros vivos estão dentro da proporção prevista pelas leis da genética de Mendel, com uma razão entre sexos igual a 1;1. Foram medidas as concentrações de galanina por meio de radioimunoensaios e por imunocitoquímica em zonas onde previamente se havia demonstrado que havia expressão da galanina a concentrações elevadas, tendo sido incluídos o cérebro, a pituitária, a medula espinal, o gânglio da raiz dorsal, o estômago, o intestino delgado e o útero. As concentrações de galanina nos animais heterozigóticos relativamente à supressão foram 50% das contraprovas de tipo natural, ao passo que as concentrações de galanina nos animais homozigóticos relativamente à supressão foram indetectáveis em todos os casos.9 clones were identified in which one allele of the galanin gene was correctly identified by homologous recombination among 209 doubly resistant colonies, determining a 4.3% labeling frequency. These nine clones were karyotypic, confirmed euploidy and injected into 3.5 day old blasts of C57 / BL / 6 mice. Germline transmission of the destroyed galanin locus was obtained from three separate clones of EE cells. The progeny genotype was determined using Southern blot analysis and PCR methodology (Fig. 4 demonstrates identical results obtained by the Southern blot analysis methodology and by PCR screening in the same litter obtained from a mating of two heterozygous animals). Mutation 7 proceeded with procreation up to homozygosity in the serotype-129 sv strain and all data presented are from the mouse with this formation. 1. The results of the analysis of the genotypes of the living unborn are within the proportion predicted by Mendel's laws of genetics, with a ratio between sexes equal to 1; 1. Galanin concentrations were measured by radioimmunoassays and by immunocytochemistry in areas where it had previously been shown that galanin was expressed at high concentrations, including the brain, pituitary, spinal cord, dorsal root ganglion, stomach , the small intestine and the uterus. The concentrations of galanin in the heterozygous animals relative to the suppression were 50% of the wild-type controls, whereas the concentrations of galanin in the homozygous animals relative to the suppression were undetectable in all cases.
Na Tabela 1 estão indicados os dados de comparação das concentrações da expressão de galanina entre murganhos de tipo natural, heterozigóticos e mutantes, em vários tecidos corporais.Comparison data of galanin expression concentrations between wild-type, heterozygous and mutant mice in various body tissues are given in Table 1.
Tabela 1Table 1
Genotipo Córtex Hipotálamo Pituitária anterior Estômago Duodeno íleo + / + 5,78±0,33 110,34±7,81 0,42±0,07 27,46±1,91 122,90±11,60 267,43+13,46 + /- 2,91±0,21 53,82±3,76 0,21±0,04 13,8±0,83 68,36±5,67 125,8717,55 ND ND ND ND ND NDGenotype Cortex Hypothalamus Anterior pituitary Stomach Duodenum ileus + / + 5.78 ± 0.33 110.34 ± 7.81 0.42 ± 0.07 27.46 ± 1.91 122.90 ± 11.60 267.43+ 13.46 +/- 2.91 ± 0.21 53.82 ± 3.76 0.21 ± 0.04 13.8 ± 0.83 68.36 ± 5.67 125.8717.55 ND ND ND ND ND ND
Todos os valores são valores médios de galanina-LI pmol/g ± SEM, excepto no caso da pituitária anterior das fêmeas, em que o valor é expresso em pmol/glângula ± SEM ND = Não detectável.All values are mean values of galanin-LI pmol / g ± SEM, except in the case of the females' anterior pituitary, where the value is expressed in pmol / gland. SEM SEM ND = Not detectable.
Verificar-se-à que a galanina não foi detectada em nenhum dos tecidos testados nos murganhos mutantes homozigóticos e que diminuiu em 50% nos murganhos mutantes heterozigóticos.It will be seen that galanin was not detected in any of the tissues tested in the homozygous mutant mice and decreased by 50% in the heterozygous mutant mice.
2. Mediu-se as aptidões regenerativas dos axónios sensoriais nos nervos ciáticos directamente pelo teste do beliscão (Danielsen, N., Kerns, J. M., Holmquist, B., Zhao, O., Lundborg, G. & Kanje, M. Brain Res. 681, 105-108, 1995). A seguir ao esmagamento dos nervos, os axónios sensoriais regeneram-se, dando origem a nervos distais e podem ser estimulados por outro beliscão subsequente dos nervos, que induz uma resposta motora reflexa abdominal. Os primeiros axónios regenerativos são localizados beliscando segmentos consecutivos do nervo num sentido que progride desde uma posição distai até uma posição proximal, até se observar um reflexo, sendo calculada a distância a partir do local de esmagamento do nervo. Num exemplo comparativo, a regeneração comprovou a existência de uma redução estatisticamente significativa da ordem de 30-40% nos animais homozigóticos comparativamente com os murganhos de tipo selvagem, aos 2, 4 e 6 dias após o esmagamento dos nervos (Fig. 5). A regeneração foi intermédia nos murganhos heterozigóticos, mas continuou a ser significativamente diferente dos animais de tipo selvagem.2. The regenerative abilities of the sensory axons in the sciatic nerves were measured directly by the pinch test (Danielsen, N., Kerns, JM, Holmquist, B., Zhao, O., Lundborg, G. & Kanje, M. Brain Res. 681, 105-108, 1995). Following the smashing of the nerves, sensory axons regenerate, giving rise to distal nerves and may be stimulated by a subsequent nerve pinch, which induces an abdominal reflex motor response. The first regenerative axons are located by pinching consecutive nerve segments in a direction that progresses from a distal position to a proximal position until a reflex is observed, and the distance from the nerve crush site is calculated. In a comparative example, regeneration has shown a statistically significant reduction of about 30-40% in homozygous animals compared to wild-type mice at 2, 4 and 6 days after nerve crushing (Fig. 5). Regeneration was intermediate in heterozygous mice, but remained significantly different from wild-type animals.
Para se testar a hipótese de uma taxa reduzida de regeneração afectar, em murganhos com falta de galanina, a 9 recuperação funcional a seguir a uma lesão por esmagamento, testou-se uma correlação de regeneração comportamental, utilizando o índice de desdobramento dos dedos das patas (Hoogeveen, J.F., Van Der Kracht, A.H., Wondergem, J., Gonzalez Gonzalez, D. & Haveman, J. Neurotoxicology, 14, 1-7, 1993) . Os roedores desdobram os dedos das suas patas traseiras quando estes entram em contacto com uma superfície sólida, o que é uma resposta que exige enervação sensorial. Assim, os dedos deixam de se desdobrar a seguir a uma axotomia, até ocorrer nova enervação dos axónios sensoriais. Mediu-se a distância de desdobramento durante 6 semanas após o esmagamento unilateral do nervo ciático direito e fez-se a comparação com a pata lateral correspondente intacta (pata esquerda). 0 desdobramento dos dedos nos murganhos de tipo natural regressou à situação normal em menos de 3 semanas após o esmagamento do nervo ciático, mas a regeneração funcional continuou incompleta ao fim de seis semanas nos murganhos mutantes (Fig. 6). 3. A menor regeneração e a menor autonomia nos murganhos com falta de galaninas podem estar eventualmente relacionadas com a morte dos neurónios a seguir à axotomia, especialmente aqueles neurónios com aptidão para a expressão normal de galanina a seguir à lesão. Num outro exemplo comparativo, para se testar a hipótese de a galanina ser essencial à sobrevivência dos neurónios durante o desenvolvimento, estudou-se a distribuição de neurónios galaninérgicos em murganhos de tipo natural e em murganhos mutantes. Uma vez que não se conseguiu visualizar os neurónios galaninérgicos nos animais mutantes ao nível das proteínas, estudou-se a expressão do ARNm, utilizando uma ribossonda específica para o sexto exão (marcada com B; ver a Fig. 3) . Para se confirmar a sobrevivência de outras populações de neurónios 10 em que há expressão da galanina, utilizou-se a ribossonda especifica do sexto exão para se visualizar os neurónios galaninérgicos no hipocampo e o núcleo paraventricular do hipotálamo de murganhos adultos de tipo natural e mutantes (Fig. 7). Não foram observadas nenhumas diferenças de expressão entre os grupos, sugerindo tal facto que o desenvolvimento neuronal foi normal nesses animais e não dependeu da galanina.To test the hypothesis that a reduced rate of regeneration affects, in mice lacking galanin, functional recovery following a crush injury, a behavioral regeneration correlation was tested using the index of finger flexion of the legs (Hoogeveen, JF, Van Der Kracht, AH, Wondergem, J., Gonzalez Gonzalez, D. & Haveman, J. Neurotoxicology, 14, 1-7, 1993). Rodents unfold the fingers of their hind paws when they come in contact with a solid surface, which is a response that requires sensory innervation. Thus, the fingers stop unfolding following an axotomy until new innervation of the sensory axons occurs. The unfolding distance was measured for 6 weeks after the unilateral crushing of the right sciatic nerve and comparison was made with the corresponding intact lateral paw (left paw). Finger splitting in wild-type mice returned to normal in less than 3 weeks after sciatic nerve crushing, but functional regeneration remained incomplete after six weeks in mutant mice (Fig. 6). 3. The lower regeneration and lower autonomy in galanin-deficient mice may possibly be related to the death of neurons following axotomy, especially those neurons capable of normal expression of galanin following injury. In another comparative example, in order to test the hypothesis that galanin is essential for survival of neurons during development, the distribution of galaninergic neurons in wild-type mice and in mutant mice was studied. Since galaninergic neurons could not be visualized in mutant animals at the protein level, mRNA expression was studied using a sixth exon specific riboprobe (labeled with B, see Fig. 3). To confirm the survival of other populations of neurons in which galanin is expressed, the sixth exon-specific riboprobe was used to visualize the galaninergic neurons in the hippocampus and the paraventricular nucleus of the hypothalamus of wild-type and mutant adult mice ( Fig. 7). No differences in expression were observed between the groups, suggesting that neuronal development was normal in these animals and did not depend on galanin.
Utilizou-se, também, a ribossonda específica do sexto exão para estudar a distribuição dos neurónios galaninérgicos do GAD duas semanas após a axotomia do nervo ciático. Observou-se um nítido aumento, por externalização, das concentrações e do número de células em que há expressão da galanina nos neurónios do DRG dos murganhos de tipo natural (Fig. 7) . No entanto, não houve nenhuma expressão nos murganhos homozigóticos com falta de galanina, sugerindo que a galanina é necessária para que tais células sobrevivam à axotomia.The sixth exon specific riboprobe was also used to study the distribution of GAN galaninergic neurons two weeks after axotomy of the sciatic nerve. There was a sharp increase in outgrowth of the concentrations and number of cells in which galanin expression is present in the DRG neurons of wild-type mice (Fig. 7). However, there was no expression in homozygous mice lacking galanin, suggesting that galanin is required for such cells to survive axotomy.
Estes resultados, referentes à regeneração e à sobrevivência das células, são particularmente significativos, na medida em que indicam que o gene da galanina é o primeiro gene a afectar a sistema nervoso periférico.These results, concerning regeneration and cell survival, are particularly significant in that they indicate that the galanin gene is the first gene to affect the peripheral nervous system.
Assim sendo, a invenção contempla à utilização de um agonista da galanina para o tratamento de neuropatias sensoriais periféricas resultantes, por exemplo, de diabetes mellitus ou de traumas (tais como os que são provocados por acidentes rodoviários). 4. Presume-se que a galanina esteja implicada na etiologia da doença de Alzheimer. A expressão da galanina no hipocampo 11 é aumentada nos neurónios colinérgicos quando há um decaimento dos níveis de acetilcolina e de colina-acetil-transferase (ChAT). A administração de galanina reduz o comportamento de amestração em diversos modelos com murganhos, sendo também verdadeira a proposição inversa, aquando da infusão de antagonistas da galanina. Mediu-se a potenciação de longo prazo (LTP) em murganhos de tipo natural e em murganhos mutantes. A LTP é um teste electrofisiológico em que nervos específicos do hipocampo são estimulados por meio de um choque eléctrico: Davies CH, Collingridge GL. J. Physiol. Lond. 1996; 496:451-470; Davies CH, Starkey SJ, Pozza MF, Collingridge GL. GABA Nature 1991; 349: 609-611. Este processo é executado in vitro utilizando secções muito finas de cérebro de animais mortos recentemente. Os resultados mostram que a LTP diminui 50% no stratum oriens no instante correspondente aos 80 minutos nos animais mutantes, comparativamente com os murganhos de tipo natural (Fig. 9 A vs C) . Pelo contrário, não foi observada nenhuma LTP quando a medição foi feita no stratum radiatum. A galanina existe com concentrações elevadas no stratum oriens, mas não existe no stratum radiatum. Até ao momento presente os dados demonstram que uma diminuição da LTP nos mutantes implica uma diminuição da memória e das faculdades cognitivas - estão a ser efectuados testes para a avaliação destas funções. Estes dados comprovam que um agonista da galanina é útil para o tratamento da doença de Alzheimer e da perda de memória associada, determinando um aumento da memária e das faculdades cognitivas.Accordingly, the invention contemplates the use of a galanin agonist for the treatment of peripheral sensory neuropathies resulting, for example, from diabetes mellitus or trauma (such as those caused by road accidents). 4. Galanin is presumed to be implicated in the etiology of Alzheimer's disease. Galanin expression in hippocampus 11 is increased in cholinergic neurons when there is a decrease in levels of acetylcholine and choline acetyl transferase (ChAT). The administration of galanin reduces the amestration behavior in several mouse models, and the inverse proposition, when infusing galanin antagonists, is also true. Long-term potentiation (LTP) was measured in wild type mice and in mutant mice. LTP is an electrophysiological test in which specific hippocampal nerves are stimulated by means of an electric shock: Davies CH, Collingridge GL. J. Physiol. Lond. 1996; 496: 451-470; Davies CH, Starkey SJ, Pozza MF, Collingridge GL. GABA Nature 1991; 349: 609-611. This process is performed in vitro using very fine brain sections from recently killed animals. The results show that LTP decreases 50% in stratum oriens at the time corresponding to 80 minutes in mutant animals, compared to wild type mice (Fig. 9A vs C). In contrast, no LTP was observed when measurement was performed on the stratum radiatum. Galanin exists with high concentrations in stratum oriens, but does not exist in stratum radiatum. To date the data demonstrate that a decrease in LTP in mutants implies a decrease in memory and cognitive faculties - tests are being performed for the evaluation of these functions. These data prove that a galanin agonist is useful for the treatment of Alzheimer's disease and associated memory loss, leading to an increase in memory and cognitive abilities.
Lisboa, 4 de Maio de 2010 12Lisbon, May 4, 2010 12
Claims (2)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GBGB9615551.0A GB9615551D0 (en) | 1996-07-24 | 1996-07-24 | Galanin |
GBGB9623869.6A GB9623869D0 (en) | 1996-11-15 | 1996-11-15 | Galanin |
Publications (1)
Publication Number | Publication Date |
---|---|
PT1342410E true PT1342410E (en) | 2010-05-11 |
Family
ID=26309752
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PT97932939T PT918455E (en) | 1996-07-24 | 1997-07-24 | USING GALANIN TO REPAIR LESOES DOS NERVOS |
PT02028584T PT1342410E (en) | 1996-07-24 | 1997-07-24 | Use of a galanin agonist in the preparation of a medicament for improving memory and other cognitive functions |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PT97932939T PT918455E (en) | 1996-07-24 | 1997-07-24 | USING GALANIN TO REPAIR LESOES DOS NERVOS |
Country Status (11)
Country | Link |
---|---|
US (1) | US20030009777A1 (en) |
EP (2) | EP0918455B1 (en) |
JP (1) | JP2000516212A (en) |
AT (2) | ATE237346T1 (en) |
AU (1) | AU3630297A (en) |
DE (2) | DE69739760D1 (en) |
DK (2) | DK1342410T3 (en) |
ES (2) | ES2340984T3 (en) |
GB (1) | GB2331301C (en) |
PT (2) | PT918455E (en) |
WO (1) | WO1998003059A1 (en) |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0819167A4 (en) | 1996-01-24 | 2002-06-12 | Synaptic Pharma Corp | Dna encoding galanin galr2 receptors and uses thereof |
US6329197B2 (en) | 1996-10-09 | 2001-12-11 | Synaptic Pharmaceutical Corporation | DNA encoding galanin GALR3 receptors and uses thereof |
US20030092042A1 (en) * | 2001-08-27 | 2003-05-15 | David Mu | Amplified oncogenes and their involvement in cancer |
WO2003070950A1 (en) * | 2002-02-22 | 2003-08-28 | Takeda Chemical Industries, Ltd. | Novel dna and use thereof |
GB0403509D0 (en) * | 2004-02-17 | 2004-03-24 | Neuro Targets Ltd | Galanin receptors and brain injury |
JP2008517930A (en) * | 2004-10-21 | 2008-05-29 | トランス テック ファーマ,インコーポレイテッド | Bissulfonamide compounds, compositions, and methods of use as agonists of GalR1 |
GB0523550D0 (en) | 2005-11-18 | 2005-12-28 | Hunter Fleming Ltd | Therapeutic uses of steroidal compounds |
AU2008207287A1 (en) * | 2007-01-19 | 2008-07-24 | Howard Florey Institute Of Experimental Physiology And Medicine | Use of galanin in a method of treating neurodegenerative diseases or conditions |
KR20100061679A (en) * | 2007-09-11 | 2010-06-08 | 몬도바이오테크 래보래토리즈 아게 | Thyrotropin releasing hormone for therapeutic applications |
EP2187946A1 (en) * | 2008-09-09 | 2010-05-26 | Mondobiotech Laboratories AG | Use of a peptide as a therapeutic agent |
ES2883838A1 (en) * | 2020-06-04 | 2021-12-09 | Univ Malaga | PREVENTION AND/OR TREATMENT OF COGNITIVE IMPAIRMENT ASSOCIATED WITH DEMENTIA SYNDROMES (Machine-translation by Google Translate, not legally binding) |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1992012997A1 (en) * | 1991-01-16 | 1992-08-06 | The General Hospital Corporation | Human galanin |
AU1462692A (en) * | 1991-02-25 | 1992-09-15 | Board Of Regents Of The University Of Washington, The | Methods for detecting galanin antagonists |
CA2105572A1 (en) * | 1991-03-06 | 1992-09-07 | Helen F. Evans | Human galanin, cdna clones encoding human galanin and a method of producing human galanin |
SE9101472D0 (en) * | 1991-05-15 | 1991-05-15 | Trion Forskning & Utveckling | GALANIN ANTAGONIST |
JPH06172387A (en) * | 1992-12-11 | 1994-06-21 | Aibaitsu Kk | Synthetic peptide derivative |
US6225282B1 (en) * | 1996-01-05 | 2001-05-01 | Genentech, Inc. | Treatment of hearing impairments |
EP0819167A4 (en) * | 1996-01-24 | 2002-06-12 | Synaptic Pharma Corp | Dna encoding galanin galr2 receptors and uses thereof |
-
1997
- 1997-07-24 AT AT97932939T patent/ATE237346T1/en not_active IP Right Cessation
- 1997-07-24 EP EP97932939A patent/EP0918455B1/en not_active Expired - Lifetime
- 1997-07-24 DK DK02028584.7T patent/DK1342410T3/en active
- 1997-07-24 GB GB9901264A patent/GB2331301C/en not_active Expired - Fee Related
- 1997-07-24 AT AT02028584T patent/ATE457125T1/en not_active IP Right Cessation
- 1997-07-24 DE DE69739760T patent/DE69739760D1/en not_active Expired - Lifetime
- 1997-07-24 DK DK97932939T patent/DK0918455T3/en active
- 1997-07-24 ES ES02028584T patent/ES2340984T3/en not_active Expired - Lifetime
- 1997-07-24 PT PT97932939T patent/PT918455E/en unknown
- 1997-07-24 WO PCT/GB1997/001991 patent/WO1998003059A1/en active IP Right Grant
- 1997-07-24 EP EP02028584A patent/EP1342410B1/en not_active Expired - Lifetime
- 1997-07-24 ES ES97932939T patent/ES2196349T3/en not_active Expired - Lifetime
- 1997-07-24 PT PT02028584T patent/PT1342410E/en unknown
- 1997-07-24 DE DE69721005T patent/DE69721005T2/en not_active Expired - Lifetime
- 1997-07-24 JP JP10506711A patent/JP2000516212A/en not_active Ceased
- 1997-07-24 US US09/230,463 patent/US20030009777A1/en not_active Abandoned
- 1997-07-24 AU AU36302/97A patent/AU3630297A/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
GB2331301C (en) | 2005-05-22 |
AU3630297A (en) | 1998-02-10 |
ES2196349T3 (en) | 2003-12-16 |
PT918455E (en) | 2003-09-30 |
EP1342410A3 (en) | 2003-12-10 |
DE69721005T2 (en) | 2004-06-09 |
EP1342410A2 (en) | 2003-09-10 |
ATE457125T1 (en) | 2010-02-15 |
ATE237346T1 (en) | 2003-05-15 |
DK0918455T3 (en) | 2003-08-04 |
EP1342410B1 (en) | 2010-02-10 |
EP0918455B1 (en) | 2003-04-16 |
US20030009777A1 (en) | 2003-01-09 |
GB2331301B (en) | 2001-02-14 |
DE69721005D1 (en) | 2003-05-22 |
DK1342410T3 (en) | 2010-05-31 |
JP2000516212A (en) | 2000-12-05 |
GB9901264D0 (en) | 1999-03-10 |
DE69739760D1 (en) | 2010-03-25 |
EP0918455A1 (en) | 1999-06-02 |
ES2340984T3 (en) | 2010-06-14 |
WO1998003059A1 (en) | 1998-01-29 |
GB2331301A (en) | 1999-05-19 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Ratcliff et al. | Production of a severe cystic fibrosis mutation in mice by gene targeting | |
Von Koch et al. | Generation of APLP2 KO mice and early postnatal lethality in APLP2/APP double KO mice | |
Evers et al. | Impairment of L-type Ca2+ channel-dependent forms of hippocampal synaptic plasticity in mice deficient in the extracellular matrix glycoprotein tenascin-C | |
Marsh et al. | Role of the Y5 neuropeptide Y receptor in feeding and obesity | |
US7795495B2 (en) | Transgenic mice for screening for inhibitors of protein aggregation and methods for making and using them | |
Bervini et al. | Mouse models of Prader–Willi syndrome: a systematic review | |
Wang et al. | Isoform‐specific knockout of FE65 leads to impaired learning and memory | |
PT1342410E (en) | Use of a galanin agonist in the preparation of a medicament for improving memory and other cognitive functions | |
ES2908669T3 (en) | Non-human animals that have a genetically modified ANGPTL8 gene | |
US5866756A (en) | Dopamine transporter knockout mice | |
US5723719A (en) | Transgenic mouse as model for diseases involving dopaminergic dysfunction | |
US6452065B2 (en) | Transgenic mouse expressing non-native wild-type and familial Alzheimer's Disease mutant presenilin 1 protein on native presenilin 1 null background | |
US6177242B1 (en) | Genomic DNA fragments containing regulatory and coding sequences for the β2-subunit of the neuronal nicotinic acetylcholine receptor and transgenic animals made using these fragments or mutated fragments | |
US6777236B1 (en) | Process for producing a neuronal host cell in vitro comprising regulatory sequences of the β2-subunit of the neuronal nicotinic acetylcholine receptor | |
US7727952B2 (en) | Methods for treating spinal and bulbar muscular atrophy using LHRH analogs | |
CN107974463B (en) | Slc6a11 gene and application of protein thereof | |
WO2006015453A1 (en) | Modified dynorphin expression in animals and identification of compounds for treatment of obesity and diabetes | |
JP3820485B2 (en) | Non-human animal that reproduces pathophysiology of bulbar spinal muscular atrophy and therapeutic agent for bulbar spinal muscular atrophy | |
Høj et al. | Possible association of growth hormone gene polymorphism with growth hormone release in calves from lines selected for high and low milk fat yield | |
US8524976B2 (en) | Iduronidase knock-out mouse | |
US20120124682A1 (en) | Dhx36 / rhau knockout mice as experimental models of muscular dystrophy | |
US20050014834A1 (en) | Selective anesthetic agents and methods of identifying the same | |
Salmen et al. | Impairment of L-type Ca2+ channel-dependent forms of hippocampal synaptic plasticity in mice deficient in the extracellular... | |
Oh | Genomic and functional analysis of vesicular inhibitory amino acid transporter during mouse embryogenesis | |
WO2002046421A2 (en) | Methods and compositions for analysis of m3 muscarinic acetylchloine receptors |